Establishment and genomic characterization of a novel retinoblastoma cell line without RB1 mutationderived from a Han Chinese donor / 上海交通大学学报（医学版）

ABSTRACT

Objective: To develop and characterize a novel retinoblastoma cell line from a Han Chinese patient. Methods: Cells were dispersed from tumor tissue harvested from an enucleated eyeball harbouring retinoblastoma under sterile conditions. The dissociated cells were cultured, purified and passaged in vitro. Morphologic and genetic analysis and detection of surface biomarkers were performed on the cell line and primary tumor by immunofluorescence, immunohistochemical staining, DNA short tandem repeat (STR) analysis, karyotype analysis, and exome sequencing. Results: This human retinoblastoma cell line was designated as SNPH-Rb-C24. It expressed neural cell adhesion molecule 1 and synaptophysin, which confirmed its neuronal derivation. DNA STR analysis showed an identical match between SNPH-Rb-C24 and primary tumor. Karyotype analysis showed complex chromosomal abnormalities in SNPH-Rb-C24, while no alteration in 13q was observed. Comparative exome sequencing identified common mutated genes and RB1+/+ in both SNPH-Rb-C24 and primary tumor. Orthotopic xenograft tumors derived from early passage cells were established. Conclusion: A human retinoblastoma cell line (SNPH-Rb-C24) derived from a Han Chinese patient with RB1+/+ retinoblastoma is developed, which retains critical biological and genomic features of the donor tumor.